Luckau Protocols

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(Tara Luckau Protocols)
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==Tara Luckau Protocols==
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Clark Laboratory
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| width="600" style="background-color: #BCED91;" align="center"| <span style="font-size:32px;"> Tara's Protocols </span>
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===NanoDrop===
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|align="right" style="background-color: #9DB68C;" |
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====Purpose====
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<span style="font-size:16px;"> [[User:Tara_K._Luckau | Tara K. Luckau's Home Page]] </span>
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The NanoDrop is used to quantify genetic material.
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# Assemble cafeteria tray
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[[User:Tara_K._Luckau/Notebook/Team_ConGen | Conservation Genetics Lab Notebook]]
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#* Kim Wipes
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#* 2µL pipette
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#* pipette tips
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#* gloves
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#* USB drive
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#* water and buffer tubes
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#* samples
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# The NanoDrop is located in North Life Sciences, room 325A
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#*
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===Agarose Gel===
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[[Luckau_Protocols | Tara's Protocols]]
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====Purpose====
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Agarose gels are used to verify the presence and estimate the size of DNA (both genomic and amplified). When loaded into a gel and subjected to an electric current, DNA fragments migrate to the positive terminal. Small DNA fragments migrate more rapidly than large ones. By running a commercially available ladder of fragments of known size, we can estimate the DNA fragment size in our sample of interest.
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To visualize the DNA after electrophoresis, a dye called Gel Red is added during the agarose-making process. The dye binds to the DNA and flouresces when exposed to UV light.
 
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====Materials To Be Familiar With====
 
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* Gel Rig, caster tray, combs
 
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* [[Image:OwlA2Large.jpg]]  [[Image:OwlA2Large_draw.jpg|300 px]]
 
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====Protocol====
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==Protocols used in Dr. Rulon Clark's laboratory==
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# Make gel using 1x TAE and agarose
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#* Use 1% for genomic DNA, 2% for amplified DNA
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{| border="1" style="margin: 1em auto 1em auto" "text-align: center"
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===Lab Processes===
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! Gel Rig Size !! 1x TAE (mL) !! 1.5% agarose (g) !! 2% agarose (g) !! Gel Red (µL) !! Sample volume (µL) !! max voltage (V)
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* [[Luckau_Protocols:NanoDrop | NanoDrop]]
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| small || 50 || _ || 1 || 5 || 4-6 || 80
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* [[Luckau_Protocols:PCR | PCR]]
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* [[Luckau_Protocols:Agarose Gel | Agarose Gel]]
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| rowspan="2"| medium || rowspan="2"| 130 || rowspan="2"| _ || rowspan="2"| 2.6 || rowspan="2"| 13 || 24-tooth comb: 6-10 || rowspan="2"| 120
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* [[Luckau_Protocols:FragmentAnalysisSubmission | Fragment Analysis Submission]]
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* [[Luckau_Protocols:ShipDryIce | Shipping with Dry Ice]]
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| 36-tooth comb: 3-5
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| large || _ || _ || _ || _ || _ || 170
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===Reagents===
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#
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* [[Luckau_Protocols:Low TE | Low TE]]
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* [[Luckau_Protocols:Tris-Cl | Tris-Cl]]
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* [[Luckau_Protocols:KCl | KCl]]
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* [[Luckau_Protocols:MgCl2 | MgCl<sub>2</sub>]]
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* [[Luckau_Protocols:TAE | TAE]]
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* [[Luckau_Protocols:1Kb Ladder | 1Kb Ladder]]
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* [[Luckau_Protocols:dNTPs | dNTPs]]
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* [[Luckau_Protocols:PCR Buffers A-H | PCR Buffers A-H]]
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* [[Luckau_Protocols:PrimerResuspension | Primer Resuspension]]
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===Software Programs===
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* [[Luckau_Protocols:GeneMarker | GeneMarker]]
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* [[Luckau_Protocols:Scoring | How I Score Frag Data]]
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* [[Luckau_Protocols:STRUCTURE | STRUCTURE]]
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===Inventory===
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* [[Luckau_Protocols:Inventory | Inventory & Catalog #s]]

Current revision

Tara's Protocols

Tara K. Luckau's Home Page

Conservation Genetics Lab Notebook

Tara's Protocols


Contents

Protocols used in Dr. Rulon Clark's laboratory

Lab Processes


Reagents


Software Programs


Inventory

Personal tools